The nervous system is a complex arrangement of neurons, their axonal projections, glia and vascular supply, yet our concepts about information processing are heavily neuron-centric. This bias results from the efficiency of standard anatomical techniques to map synaptic connections between neurons relative to other physical interactions among neural and non-neural cells. Spatial organization of neuronal groups and glia networks is indicated in many studies, but spatial organization and interactions have not been studied with an integrated approach. We will employ a new technology, serial block-face scanning electron microscopy, to reconstruct, at high resolution, all cellular elements of large tissue volumes to identify new principles in nervous system cellular organization. We will tap into human visual-cognitive capabilities to understand spatial order by developing procedures, implemented in a software user interface, to display combinations of cellular elements and vascular structures in 3-D virtual reality environments. Identification of structural features that define 3D organization of tissue will facilitate investigation of large-scale tissue volumes by othr investigators as high-throughput, high resolution studies of brain structure become more common. PUBLIC HEALTH RELEVANCE: The goal of this project is to reveal new principles of tissue organization in the brain. We expect that our experimental results will provide a basis for new metrics to evaluate brain pathology and therapeutic strategies.